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X-Ray Crystallographic Studies of Glycerol-3-Phosphate Cytidylyltransferase from Staphylococcus Aureus / The Structure of Glycerol-3-Phosphate Cytidylyltransferase from Staphylococcus Aureus

Glycerol-3-phosphate cytidylyltransferase from 𝘚𝘡𝘒𝘱𝘩𝘺𝘭𝘰𝘀𝘰𝘀𝘀𝘢𝘴 𝘒𝘢𝘳𝘦𝘢𝘴 complexed with CTP (TarDₛₐ-CTP) was crystallized by the hanging drop-vapor diffusion method at 22Β°C. Determination of crystallization condition included examination of the amount of precipitant, investigation of the effects of small molecules, and alteration of the rate of diffusion. With these three optimization steps, crystals suitable for x-ray diffraction study were produced. During data processing, TarDₛₐ-CTP was determined to belong to the space group P3₁21, with unit-cell dimensions a=b=92.2 and c=156.1Γ…. The crystal structure of TarDₛₐ-CTP was solved to 3.0Γ… by molecular replacement, using TagD from π˜‰π˜’π˜€π˜ͺ𝘭𝘭𝘢𝘴 𝘴𝘢𝘣𝘡π˜ͺ𝘭π˜ͺ𝘴 as a search model. Unlike the search model, TarDₛₐ appears as a tetramer in the asymmetric unit. This result also confirms the gel-filtration and ultracentrifugation studies that were done previously. Although TarDₛₐ crystals were grown in the presence of CTP, the crystal structure does not reveal convincing data for the location and position of this co-factor. However, the data suggests a possible location for CTP in one of the four subunits in an orientation that differs from that of TagD_Bβ‚›. Unfortunately, the resolution of this data set at 3.0Γ… is not high enough to corroborate this finding. / Thesis / Master of Science (MS)

Identiferoai:union.ndltd.org:mcmaster.ca/oai:macsphere.mcmaster.ca:11375/22976
Date January 2002
CreatorsYim, Veronica
ContributorsBerghuis, A. M., Biochemistry
Source SetsMcMaster University
LanguageEnglish
Detected LanguageEnglish
TypeThesis

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